冯微宏;韩毅;王雅静;肖勇;

• 1:无锡市疾病预防控制中心微生物检验部

摘要(Abstract)：

对引起一起无菌性脑膜炎的埃可病毒30型（（Echovirus 30,E30）毒株进行全基因组测序，并分析其遗传变异和分子进化特征。提取病毒RNA，荧光RT-PCR确定为肠道病毒，再扩增其VP1区，测序确定为E30，设计针对E30全基因组的引物，RT-PCR扩增和序列测定获得全基因序列。利用DNAMAN 9.0、MEGA X、RDP 5和SimPlot 3.5.1软件分析全基因序列。E30无锡株基因组全长7 425 bp个核苷酸（nt）,5′端和3′端分别为743 nt和97 nt的UTR，二个UTR之间为一个6 585 nt长的开放阅读框，编码一个含2 195个氨基酸（aa）的多聚蛋白。与GenBank中基因组序列比对，最同源的是毒株USA/2017/CA-RGDS-1048 （基因登录号：MN153801），核苷酸同源性为97.5%，氨基酸同源性为99.1%。VP1基因进化树分析显示，E30无锡株属于h型，并可归类于h型下分的基因簇GroupⅣ。种系进化分析和同源性分析提示E30无锡株可上溯到中国江苏毒株FDJS03毒株。分析还发现E30无锡株在非结构区存在重组，重组序列可能来自E30毒株（KY888274）和E18毒株（MN815813）。E30无锡株归类为h型E30，它是一个包含了E30毒株和E18毒株序列的重组病毒。

关键词(KeyWords)： 埃可病毒30型;全基因组;序列分析;重组

Abstract:

Keywords:

基金项目(Foundation):

作者(Authors): 冯微宏;韩毅;王雅静;肖勇;

DOI: 10.13242/j.cnki.bingduxuebao.004254

参考文献(References)：

• [1] Marjom?ki V, Kalander K, Hellman M, Permi P.Enteroviruses and coronaviruses:similarities and therapeutic targets[J]. Expert Opin Ther Targets,2021, 25(6):479-489.
• [2] Sun Y, Miao Z, Yan J, Gong L, Chen Y, Chen Y,Mao H, Zhang Y. Sero-molecular epidemiology of enterovirus-associated encephalitis in Zhejiang province,China, from 2014 to 2017[J]. Int J Infect Dis, 2019,79:58-64.
• [3] Prill M M, Dahl R M, Midgley C M, Chern S W, Lu X, Feikin D R, Sakthivel S K, Nix W A, Watson J T,Gerber S I, Oberste M S. Severe respiratory illness associated with rhinovirus during the enterovirus D68outbreak in the United States, August 2014-November2014[J]. Clin Infect Dis, 2018, 66(10):1528-1534.
• [4] Rohll J B, Moon D H, Evans D J, Almond J W. The 3'untranslated region of picornavirus RNA:features required for efficient genome replication[J]. J Virol,1995, 69(12):7835-7844.
• [5] Bergamini G, Preiss T, Hentze M W. Picornavirus IRESes and the poly(A)tail jointly promote capindependent translation in a mammalian cell-free system[J]. Rna, 2000, 6(12):1781-1790.
• [6] Zhao Y N, Jiang Q W, Jiang R J, Chen L, Perlin D S.Echovirus 30, Jiangsu province, China[J]. Emerg Infect Dis, 2005, 11(4):562-567.
• [7] Yang X H, Yan Y S, Weng Y W, He A H, Zhang H R, Chen W, Zhou Y. Molecular epidemiology of echovirus 30 in Fujian, China between 2001 and 2011[J]. J Med Virol, 2013, 85(4):696-702.
• [8] Xiao H, Guan D, Chen R, Chen P, Monagin C, Li W,Su J, Ma C, Zhang W, Ke C. Molecular characterization of echovirus 30-associated outbreak of aseptic meningitis in Guangdong in 2012[J]. Virol J,2013, 10:263.
• [9] Zheng S, Ye H, Yan J, Xie G, Cui D, Yu F, Wang Y, Yang X, Zhou F, Zhang Y, Tian X, Chen Y.Laboratory diagnosis and genetic analysis of a family clustering outbreak of aseptic meningitis due to echovirus30[J]. Pathog Glob Health, 2016, 110(6):233-237.
• [10]Oberste M S, Maher K, Williams A J, DybdahlSissoko N, Brown B A, Gookin M S, Pe?aranda S,Mishrik N, Uddin M, Pallansch M A. Species-specific RT-PCR amplification of human enteroviruses:a tool for rapid species identification of uncharacterized enteroviruses[J]. J Gen Virol, 2006, 87(Pt 1):119-128.
• [11]Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5:molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Mol Biol Evol, 2011, 28(10):2731-2739.
• [12]Felsenstein J. Confidence limits on phylogenies:an approach using the bootstrap[J]. Evolution, 1985, 39(4):783-791.
• [13]Martin D P, Murrell B, Golden M, Khoosal A, Muhire B. RDP4:Detection and analysis of recombination patterns in virus genomes[J]. Virus Evol, 2015, 1(1):vev003.
• [14]Lole K S, Bollinger R C, Paranjape R S, Gadkari D,Kulkarni S S, Novak N G, Ingersoll R, Sheppard H W, Ray S C. Full-length human immunodeficiency virus type 1 genomes from subtype C-infected seroconverters in India, with evidence of intersubtype recombination[J]. J Virol, 1999, 73(1):152-160.
• [15]Bailly J L, Mirand A, Henquell C, Archimbaud C,Chambon M, CharbonnéF, TraoréO, Peigue-Lafeuille H. Phylogeography of circulating populations of human echovirus 30 over 50 years:nucleotide polymorphism and signature of purifying selection in the VP1 capsid protein gene[J]. Infect Genet Evol, 2009, 9(4):699-708.
• [16]Tapparel C, Siegrist F, Petty T J, Kaiser L.Picornavirus and enterovirus diversity with associated human diseases[J]. Infect Genet Evol, 2013, 14:282-293.
• [17]Adams M J, Lefkowitz E J, King A M Q, Harrach B,Harrison R L, Knowles N J, Kropinski A M, Krupovic M, Kuhn J H, Mushegian A R, Nibert M,Sabanadzovic S, Sanfa?on H, Siddell S G, Simmonds P, Varsani A, Zerbini F M, Gorbalenya A E, Davison A J. Changes to taxonomy and the international code of virus classification and nomenclature ratified by the international committee on taxonomy of viruses(2017)[J]. Arch Virol, 2017, 162(8):2505-2538.
• [18]Oberste M S, Maher K, Kilpatrick D R, Pallansch M A. Molecular evolution of the human enteroviruses:correlation of serotype with VP1 sequence and application to picornavirus classification[J]. J Virol,1999, 73(3):1941-1948.
• [19]Chen P, Lin X, Liu G, Wang S, Song L, Tao Z, Xu A. Analysis of enterovirus types in patients with symptoms of aseptic meningitis in 2014 in Shandong,China[J]. Virology, 2018, 516:196-201.
• [20]Bailly J L, Béguet A, Chambon M, Henquell C, Peigue-Lafeuille H. Nosocomial transmission of echovirus 30:molecular evidence by phylogenetic analysis of the VP1encoding sequence[J]. J Clin Microbiol, 2000, 38(8):2889-2892.
• [21]Holmes C W, Koo S S, Osman H, Wilson S, Xerry J,Gallimore C I, Allen D J, Tang J W. Predominance of enterovirus B and echovirus 30 as cause of viral meningitis in a UK population[J]. J Clin Virol, 2016,81:90-93.
• [22]Chen J, Han Z, Wu H, Xu W, Yu D, Zhang Y. A large-scale outbreak of echovirus 30 in Gansu Province of China in 2015 and its phylodynamic characterization[J].Front Microbiol, 2020, 11:1137.
• [23]Zhao Y N, Perlin D S, Park S, Jiang R J, Chen L,Chen Y, Gardiner R, Jiang Q W. FDJS03 isolates causing an outbreak of aseptic meningitis in China that evolved from a distinct echovirus 30 lineage imported from countries of the commonwealth of independent states[J]. J Clin Microbiol, 2006, 44(11):4142-4148.
• [24]Gavrilin G V, Cherkasova E A, Lipskaya G Y, Kew O M, Agol V I. Evolution of circulating wild poliovirus and of vaccine-derived poliovirus in an immunodeficient patient:a unifying model[J]. J Virol, 2000, 74(16):7381-7390.
• [25]Dilcher M, Howard J C, Dalton S C, Anderson T,Clinghan R T, Werno A M. Clinical, laboratory, and molecular epidemiology of an outbreak of aseptic meningitis due to a triple-recombinant echovirus in Ashburton, New Zealand[J]. Viruses, 2022, 14(4).
• [26]Oberste M S, Maher K, Pallansch M A. Evidence for frequent recombination within species human enterovirus B based on complete genomic sequences of all thirtyseven serotypes[J]. J Virol, 2004, 78(2):855-867.
• [27]Lukashev A N, Lashkevich V A, Ivanova O E,Koroleva G A, Hinkkanen A E, Ilonen J.Recombination in circulating Human enterovirus B:independent evolution of structural and non-structural genome regions[J]. J Gen Virol, 2005, 86(Pt 12):3281-3290.
• [28]Tian X, Han Z, He Y, Sun Q, Wang W, Xu W, Li H, Zhang Y. Temporal phylogeny and molecular characterization of echovirus 30 associated with aseptic meningitis outbreaks in China[J]. Virol J, 2021, 18(1):118.
• [29]Lukashev A N, Shumilina E Y, Belalov I S, Ivanova O E, Eremeeva T P, Reznik V I, Trotsenko O E,Drexler J F, Drosten C. Recombination strategies and evolutionary dynamics of the human enterovirus A global gene pool[J]. J Gen Virol, 2014, 95(Pt 4):868-873.
• [30]Chen P, Song Z, Qi Y, Feng X, Xu N, Sun Y, Wu X, Yao X, Mao Q, Li X, Dong W, Wan X, Huang N,Shen X, Liang Z, Li W. Molecular determinants of enterovirus 71 viral entry:cleft around GLN-172 on VP1 protein interacts with variable region on scavenge receptor B 2[J]. J Biol Chem, 2012, 287(9):6406-6420.